GSM4711923: FOXC1-LSCs rep2; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
FOXC1
Cell type
Cell type Class
Others
Cell type
Limbal stem cells
NA
NA
Attributes by original data submitter
Sample
source_name
limbal stem cells
cell type
primary limbal stem cells
genotype
WT
shRNA
none
chip antibody
FOXC1 (Bethyl Laboratories, A303-519A)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
The RNeasy Mini Kit (Qiagen) was used to isolate total RNA from cells. For ChIP-seq, chromatin was fixed with 1% formaldehyde for 10 min and sheared in a sonification buffer. Fragmented DNA was incubated with primary antibodies overnight at 4℃ and then with Protein A/G Dynabeads for 1 h. The beads were collected and washed sequentially with the following solutions: high-salt buffer, low-salt wash buffer, and TE buffer. Elution of the immunoprecipitated protein/DNA complexes was performed using elution buffer at 65℃ for 4 h. Next, the protein/DNA complexes were incubated with proteinase K (Invitrogen) and RNase A (Invitrogen) for 1 h. De-crosslinked DNA was purified using the PCR Purification Kit (Qiagen). For ATAC-seq, in total, 50,000 cells were digested, collected, and lysed in ice-cold lysis buffer for 5 min. Then, Tn5 transposase reactions (Vazyme Biotech, TD501) were performed using the TruePrep DNA Library Prep Kit (Vazyme Biotech). RNA libraries were prepared for sequencing using NEBNext RNA first and second Strand Synthesis Module (NEB, USA) and KAPA Library Preparation Kit (KAPA Biosystem). The KAPA Hyper Prep Kit (Kapa Biosystems, KK8502) was used to construct ChIP-seq libraries for Illumina PE150 sequencing. For ATAC-seq library, the transposed DNA fragments were amplified and collected using the TruePrep DNA Library Prep Kit (Vazyme Biotech).